This proposal seeks to use chemical genomics to identify small-molecule compounds that modulate peroxisome biogenesis and function. Peroxisomes are major sites of fatty-acid oxidation and the only site of very long-chain fatty-acid metabolism. Ectopic fatty-acid deposition in non-adipose tissue leads to lipotoxicity, which is associated with numerous disease conditions. Increasing fatty-acid oxidation capacity in cells and shifting fatty- acid metabolism towards oxidation may prevent lipotoxicity and could be therapeutic. In addition, peroxisomes are required for the synthesis of specialized phospholipids, sphingolipids, that are required in myelin and when absent result in neurodegenerative diseases. In fact, multiple human diseases ranging from neurodegeneration to metabolic abnormalities result from the absence of functional peroxisomes. We are developing a novel high throughput screening (HTS) assay using human liver cells expressing a fluorescent peroxisome reporter to monitor peroxisome biogenesis in real time. Compounds identified our screen could be the first step towards developing PPAR independent potential therapeutics for diseases ranging from Type 2 diabetes to neurodegenerative conditions. PUBLIC HEALTH RELEVANCE: This proposal utilizes a novel high throughput screening (HTS) assay to identify novel PPAR independent regulators of peroxisome biogenesis and function. Due to peroxisomes required function for sphingolipid synthesis, which are required for myelination, decreased peroxisome function results in neurodegenerative disease (e.g. Zellwegers syndrome). The compounds identified from this proposal are a first step towards developing potential therapeutics for neurodegenerative disease, metabolic syndrome, and Type 2 diabetes.